Trophic Transfer of Cd, Cu, Pb, Zn, P and Se in Dutch Storage Water Reservoirs.

Heavy metals are naturally omnipresent in aquatic systems. Excess amounts of heavy metals can accumulate in organisms of pollution impacted systems and transfer across a food web. Analysing the food web structure and metal contents of the organisms can help unravel the pathways of biomagnification or biodilution and gain insight in trophic linkages. We measured heavy metals and other elements in mussel bank detritus and organisms of the Biesbosch reservoirs (the Netherlands) and linked those to stable isotopic signatures. The heavy metal contents (cadmium, copper, lead, and zinc) were often lowest in benthivorous, omnivorous and piscivorous species (mainly fish); whereas, phosphorus contents were lower in the autotrophs. Mussel bank detritus contained the highest amounts of heavy metals. The heavy metals were negatively correlated with δ15N values. For selenium no clear trend was observed. Furthermore, there was a negative correlation between fish length and some heavy metals. Based on all 20 analysed elemental contents, similarities between species became apparent, related to niche or habitat. This study confirms that elemental contents of species can differ between feeding guilds and/or species, which can be attributed to metabolic and physiological processes. The organisms in higher trophic levels have adaptations preventing metal accumulation, resulting in lower contents. Within the fish species biodilution occurs, as most metal contents were lowest in bigger fish. Overall, the metals did not seem to biomagnify, but biodilute in the food web. Metal analyses combined with isotopic signatures could thus provide insights in metal transfer and possible trophic linkages within a system.

Polishing wastewater effluent using plants: floating plants perform better than submerged plants in both nutrient removal and reduction of greenhouse gas emission.

While research on aquatic plants used in treatment wetlands is abundant, little is known about the use of plants in hydroponic ecological wastewater treatment, and its simultaneous effect on greenhouse gas (GHG) emissions. Here, we assess the effectiveness of floating and submerged plants in removing nutrients and preventing GHG emissions from wastewater effluent. We grew two species of floating plants, Azolla filiculoides and Lemna minor, and two species of submerged plants, Ceratophyllum demersum and Callitriche platycarpa, on a batch of domestic wastewater effluent without any solid substrate. In these systems, we monitored nitrogen and phosphorus removal and fluxes of CO2, CH4 and N2O, for 2 weeks. In general, floating plants produced the most biomass, whereas submerged plants were rapidly overgrown by filamentous algae. Floating plants removed nutrients most efficiently; both floating species removed 100% of the phosphate while Lemna also removed 97-100% of the inorganic nitrogen, as opposed to a removal of 81-88% in submerged plants with algae treatments. Moreover, aquaria covered by floating plants had roughly three times higher GHG uptake than the treatments with submerged plants or controls without plants. Thus, effluent polishing by floating plants can be a promising avenue for climate-smart wastewater polishing.

Overcoming establishment thresholds for peat mosses in human-made bog pools.

Globally, peatlands have been affected by drainage and peat extraction, with adverse effects on their functioning and services. To restore peat-forming vegetation, drained bogs are being rewetted on a large scale. Although this practice results in higher groundwater levels, unfortunately it often creates deep lakes in parts where peat was extracted to greater depths than the surroundings. Revegetation of these deeper waters by peat mosses appears to be challenging due to strong abiotic feedbacks that keep these systems in an undesired bare state. In this study, we theoretically explore if a floating peat mat and an open human-made bog lake can be considered two alternative stable states using a simple model, and experimentally test in the field whether stable states are present, and whether a state shift can be accomplished using floating biodegradable structures that mimic buoyant peat. We transplanted two peat moss species into these structures (pioneer sp. Sphagnum cuspidatum and later-successional sp. S. palustre) with and without additional organic substrate. Our model suggests that these open human-made bog lakes and floating peat mats can indeed be regarded as alternative stable states. Natural recovery by spontaneous peat moss growth, i.e., a state shift from open water to floating mats, is only possible when the water table is sufficiently shallow to avoid light limitation (<0.29 m at our site). Our experiment revealed that alternative stable states are present and that the floating structures facilitated the growth of pioneer S. cuspidatum and vascular plants. Organic substrate addition particularly facilitated vascular plant growth, which correlated to higher moss height. The structures remained too wet for the late-successional species S. palustre. We conclude that open water and floating peat mats in human-made bog lakes can be considered two alternative stable states, and that temporary floating establishment structures can induce a state shift from the open water state to peat-forming vegetation state. These findings imply that for successful restoration, there is a clear water depth threshold to enable peat moss growth and there is no need for addition of large amounts of donor-peat substrate. Correct species selection for restoration is crucial for success.

Foundation species' overlap enhances biodiversity and multifunctionality from the patch to landscape scale in southeastern United States salt marshes.

Although there is mounting evidence that biodiversity is an important and widespread driver of ecosystem multifunctionality, much of this research has focused on small-scale biodiversity manipulations. Hence, which mechanisms maintain patches of enhanced biodiversity in natural systems and if these patches elevate ecosystem multifunctionality at both local and landscape scales remain outstanding questions. In a 17 month experiment conducted within southeastern United States salt marshes, we found that patches of enhanced biodiversity and multifunctionality arise only where habitat-forming foundation species overlap--i.e. where aggregations of ribbed mussels (Geukensia demissa) form around cordgrass (Spartina alterniflora) stems. By empirically scaling up our experimental results to the marsh platform at 12 sites, we further show that mussels--despite covering only approximately 1% of the marsh surface--strongly enhance five distinct ecosystem functions, including decomposition, primary production and water infiltration rate, at the landscape scale. Thus, mussels create conditions that support the co-occurrence of high densities of functionally distinct organisms within cordgrass and, in doing so, elevate salt marsh multifunctionality from the patch to landscape scale. Collectively, these findings suggest that patterns in foundation species' overlap drive variation in biodiversity and ecosystem functioning within and across natural ecosystems.We therefore argue that foundation species should be integrated in our conceptual understanding of forces that moderate biodiversity--ecosystem functioning relationships, approaches for conserving species diversity and strategies to improve the multifunctionality of degraded ecosystems.

Seagrasses are negatively affected by organic matter loading and Arenicola marina activity in a laboratory experiment.

When two ecosystem engineers share the same natural environment, the outcome of their interaction will be unclear if they have contrasting habitat-modifying effects (e.g., sediment stabilization vs. sediment destabilization). The outcome of the interaction may depend on local environmental conditions such as season or sediment type, which may affect the extent and type of habitat modification by the ecosystem engineers involved. We mechanistically studied the interaction between the sediment-stabilizing seagrass Zostera noltii and the bioturbating and sediment-destabilizing lugworm Arenicola marina, which sometimes co-occur for prolonged periods. We investigated (1) if the negative sediment destabilization effect of A. marina on Z. noltii might be counteracted by positive biogeochemical effects of bioirrigation (burrow flushing) by A. marina in sulfide-rich sediments, and (2) if previously observed nutrient release by A. marina bioirrigation could affect seagrasses. We tested the individual and combined effects of A. marina presence and high porewater sulfide concentrations (induced by organic matter addition) on seagrass biomass in a full factorial lab experiment. Contrary to our expectations, we did not find an effect of A. marina on porewater sulfide concentrations. A. marina activities affected the seagrass physically as well as by pumping nutrients, mainly ammonium and phosphate, from the porewater to the surface water, which promoted epiphyte growth on seagrass leaves in our experimental set-up. We conclude that A. marina bioirrigation did not alleviate sulfide stress to seagrasses. Instead, we found synergistic negative effects of the presence of A. marina and high sediment sulfide levels on seagrass biomass.

Lower nitrate leaching from dairy cattle slurry compared to synthetic fertilizer calcium ammonium nitrate applied to grassland.

Nitrate leaching from agriculture can be reduced by the choice of fertilizer and a proper timing of its application. For permanent grassland grown under temperate conditions, nitrate leaching was hypothesized to be lower from dairy cattle slurry (CS) compared to synthetic fertilizer calcium ammonium nitrate (CAN), based on differences in chemical composition, consequential effects on nitrogen (N) conversion processes in soil, and resulting differences in synchronization of (nitrate) N availability and plant N uptake. We tested the hypothesis in a two-year field experiment on cut grassland on a leaching-sensitive sandy soil, fertilized each year with 320 kg ha-1 of plant-available N from either 100% top-dressed CAN or a combination of 40% from CAN and 60% from sod-injected CS, and measured effects on grass herbage yield, herbage N uptake, and nitrate concentration in pore water at 1.0 m depth. Our results show a comparable level of herbage N uptake for both treatments, allowing for a proper comparison of nitrate leaching at a similar level of plant-available N. Average nitrate concentration in pore water in the main leaching period (over winter) was after the first (dry) growing season 44% lower for CS + CAN (41 mg l-1) compared to CAN only (73 mg l-1), and after the second (wet) growing season 35% lower for CS + CAN (32 mg l-1) compared to CAN only (49 mg l-1). Nitrogen application increased nitrate concentration at 1.0 m depth not only in winter but also in the growing season. We conclude that for permanent grasslands in temperate regions, nitrate leaching from timely applied CS may be considerably lower than from CAN, which is different from previous assumptions.

Wastewater-effluent discharge and incomplete denitrification drive riverine CO2, CH4 and N2O emissions.

Rivers are well-known sources of the greenhouse gasses (GHG) carbon dioxide (CO2), methane (CH4) and nitrous oxide (N2O). These emissions from rivers can increase because of anthropogenic activities, such as agricultural fertilizer input or the discharge of treated wastewater, as these often contain elevated nutrient concentrations. Yet, the specific effects of wastewater effluent discharge on river GHG emissions remain poorly understood. Here, we studied two lowland rivers which both receive municipal wastewater effluent: river Linge and river Kromme Rijn. Dissolved concentrations and fluxes of CH4, N2O and CO2 were measured upstream, downstream and at discharge locations, alongside water column properties and sediment composition. Microbial communities in the sediment and water column were analysed using 16S rRNA gene sequencing. In general, observed GHG emissions from Linge and Kromme Rijn were comparable to eutrophic rivers in urban and agricultural environments. CO2 emissions peaked at most discharge locations, likely resulting from dissolved CO2 present in the effluent. CH4 emission was highest 2 km downstream, suggesting biological production by methanogenic activity stimulated by the effluents' carbon and nutrient supply. Dissolved N2O concentrations were strongly related to NO3- content of the water column which points towards incomplete riverine denitrification. Notably, methanogenic archaea were more abundant downstream of effluent discharge locations. However, overall microbial community composition remained relatively unaffected in both rivers. In conclusion, we demonstrate a clear link between wastewater effluent discharge and enhanced downstream GHG emission of two rivers. Mitigating the impact of wastewater effluent on receiving rivers will be crucial to reduce riverine GHG contributions.

The threat of groundwater pollution for petrifying springs; defining nutrient threshold values for an endangered bryophyte dominated habitat.

Eutrophication by human activities is increasingly affecting ecosystem functioning and plant community composition. So far, studies mainly focus on the effects of atmospheric nitrogen deposition, surface water eutrophication or soil nutrient accumulation. Groundwater pollution of spring habitats, however, has received much less attention, although numerous papers report groundwater nutrient enrichment worldwide. This study presents a survey on groundwater pollution (with emphasis on nitrate and phosphate) and bryophyte composition in 51 ambient petrifying springs in 5 NW European countries, which were compared to published data from 173 other sites in 11 European countries. The reviewed dataset covers a broad range of unpolluted to heavily polluted springs with nitrate concentrations between 0.7 and 3227 µmol l-1. Most petrifying springs in the rural lowlands of NW Europe were found to have elevated concentrations of nitrate and phosphate with the most polluted springs occurring in The Netherlands. The cover of individual characteristic bryophyte species significantly correlates with groundwater nutrient concentrations indicating that nutrient pollution of spring waters affects bryophyte composition. Palustriella commutata, Eucladium verticillatum and Brachythecium rivulare prefer unpolluted petrifying springs whereas Cratoneuron filicinum and Pellia endiviifolia show a much broader tolerance to groundwater pollution. In order to sustain at least the basic conditions for the typical bryophyte composition of petrifying springs habitats, threshold values of 288 µmol (18 mg l-1) NO3- l-1 and 0.42 µmol (0.04 mg l-1) ortho-PO43- l-1 were defined. Data analysis of the spring water composition indicates that the main source for nutrient and nutrient induced base cation enrichment are nitrate losses from intensively used agricultural fields. The anthropogenically induced but regionally different chemical processes in subsoil and aquifers can result in different levels of nutrient pollution in springs. Further regulations for nitrate and phosphate application are required to conserve and restore groundwater fed ecosystems in Europe.

Addition of iron does not ameliorate sulfide toxicity by sargassum influx to mangroves but dampens methane and nitrous oxide emissions.

Sargassum spp. strandings in the tropical Atlantic harm local ecosystems due to toxic sulfide levels. We conducted a mesocosm experiment to test the efficacy of iron(III) (hydr)oxides in (a) mitigating sulfide toxicity in mangroves resulting from Sargassum and (b) reducing potentially enhanced greenhouse gas emissions. Our results show that iron addition failed to prevent mangrove mortality caused by highly toxic sulfide concentrations, which reached up to 15,000 µmol l-1 in 14 days; timely removal may potentially prevent mangrove death. Sargassum-impacted mesocosms significantly increased methane, nitrous oxide, and carbon dioxide emissions, producing approximately 1 g CO2-equivalents m-2 h-1 during daylight hours, thereby shifting mangroves from sinks to sources of greenhouse gasses. However, iron addition decreased methane emissions by 62 % and nitrous oxide emissions by 57 %. This research reveals that Sargassum strandings have multiple adverse effects related to chemical and ecological dynamics in mangrove ecosystems, including greenhouse gas emissions.

Sludge degradation, nutrient removal and reduction of greenhouse gas emission by a Chironomus-Azolla wastewater treatment cascade.

Wastewater treatment plants (WWTPs) are a point source of nutrients, emit greenhouse gases (GHGs), and produce large volumes of excess sludge. The use of aquatic organisms may be an alternative to the technical post-treatment of WWTP effluent, as they play an important role in nutrient dynamics and carbon balance in natural ecosystems. The aim of this study was therefore to assess the performance of an experimental wastewater-treatment cascade of bioturbating macroinvertebrates and floating plants in terms of sludge degradation, nutrient removal and lowering GHG emission. To this end, a full-factorial experiment was designed, using a recirculating cascade with a WWTP sludge compartment with or without bioturbating Chironomus riparius larvae, and an effluent container with or without the floating plant Azolla filiculoides, resulting in four treatments. To calculate the nitrogen (N), phosphorus (P) and carbon (C) mass balance of this system, the N, P and C concentrations in the effluent, biomass production, and sludge degradation, as well as the N, P and C content of all compartments in the cascade were measured during the 26-day experiment. The presence of Chironomus led to an increased sludge degradation of 44% compared to 25% in the control, a 1.4 times decreased transport of P from the sludge and a 2.4 times increased transport of N out of the sludge, either into Chironomus biomass or into the water column. Furthermore, Chironomus activity decreased methane emissions by 92%. The presence of Azolla resulted in a 15% lower P concentration in the effluent than in the control treatment, and a CO2 uptake of 1.13 kg ha-1 day-1. These additive effects of Chironomus and Azolla resulted in an almost two times higher sludge degradation, and an almost two times lower P concentration in the effluent. This is the first study that shows that a bio-based cascade can strongly reduce GHG and P emissions simultaneously during the combined polishing of wastewater sludge and effluent, benefitting from the additive effects of the presence of both macrophytes and invertebrates. In addition to the microbial based treatment steps already employed on WWTPs, the integration of higher organisms in the treatment process expands the WWTP based ecosystem and allows for the inclusion of macroinvertebrate and macrophyte mediated processes. Applying macroinvertebrate-plant cascades may therefore be a promising tool to tackle the present and future challenges of WWTPs.

Early stakeholder involvement using Group Model Building to identify ecological research questions and nature management options.

Many tropical coastal ecosystems face human pressures related to tourism, land or sea use. We developed a practical procedure to involve stakeholders in an early stage of an ecological research project to map the Social-Ecological System (SES) in our case study Lac Bay, Bonaire island, as well as to identify and prioritize ecological research questions and nature management options in relation to a recent new threat: massive sargassum landings. In our procedure we used the Group Model Building methodology for identifying drivers, key variables and feedback loops in this SES. The underlying mechanisms of driving feedbacks were revealed and shared during these sessions. We identified and prioritized urgent ecological research questions for the conservation of seagrass beds and mangrove forests, and practical measures for nature management in Lac Bay. Both were used in follow-up scientific research and nature management plans, illustrating the applicability of our procedure for early science-stakeholder interaction.

CO2, CH4, and N2O emissions from dredged material exposed to drying and zeolite addition under field and laboratory conditions.

Dredging, the removal of sediment from water courses, is generally conducted to maintain their navigability and to improve water quality. Recent studies indicate that dredging can significantly reduce aquatic greenhouse gas (GHG) emissions. These studies, however, do not consider the potential emission from the dredged material (sludge) in the depot. In addition, it is unknown if and how GHG emissions from sludge depots can be reduced. Here we present spatiotemporal variations of carbon dioxide (CO2), methane (CH4), and nitrous oxide (N2O) fluxes, as well as environmental variables from a sludge depot located in the Netherlands. Measurements were conducted monthly from the time the depot was filled until the sludge was dry and the depot was abolished. We also experimentally assessed the GHG mitigation potential of 1) keeping the sludge permanently inundated, and 2) the addition of different amounts of zeolite to increase sludge nitrogen binding capacity to reduce N2O emissions. In the depot and in the laboratory, a decrease in moisture content coincided with increased CO2 and N2O emissions while CH4 emissions decreased. We observed that permanent inundation reduced emissions (∼4 times less CO2-eq than in drying sludge). Adding zeolite lowered N2O fluxes from permanently inundated sludge but did not reduce total GHG emissions. During the depot's operational period, average CO2, CH4, and N2O fluxes were 5078, 27, and 5 mg m-2 d-1, respectively. GHG emissions from drying sludge occurred mainly in the form of CO2 (73% of the total CO2-eq emissions), with average GHG emission rates comparable to those reported for ditches and ponds. We estimate that approximately 14 tons of CO2-eq were emitted from the 0.011 km2 depot, which contained ∼20,000 m3 of sludge, during its entire operational period, and we argue that more studies are needed, considering different sludge origins, to expand our understanding of sludge depots.

Nitrate-dependent anaerobic methane oxidation (N-DAMO) as a bioremediation strategy for waters affected by agricultural runoff.

Agricultural drainage ditches are subjected to high anthropogenic nitrogen input, leading to eutrophication and greenhouse gas emissions. Nitrate-dependent anaerobic methane oxidation (N-DAMO) could be a promising remediation strategy to remove methane (CH4) and nitrate (NO3-) simultaneously. Therefore, we aimed to evaluate the potential of N-DAMO to remove excess NO3- and decrease CH4 release from agricultural drainage ditches. Microcosm experiments were conducted using sediment and surface water collected from three different sites: a sandy-clay ditch (SCD), a freshwater-fed peatland ditch (FPD), and a brackish peatland ditch (BPD). The microcosms were inoculated with an N-DAMO enrichment culture dominated by Candidatus Methanoperedens and Candidatus Methylomirabilis and supplemented with 13CH4 and 15NO3-. A significant decrease in CH4 and NO3- concentration was only observed in the BPD sediment. In freshwater sediments (FPD and SCD), the effect of N-DAMO inoculation on CH4 and NO3- removal was negligible, likely because N-DAMO microorganisms were outcompeted by heterotrophic denitrifiers consuming NO3- much faster. Overall, our results suggest that bioaugmentation with N-DAMO might be a potential strategy for decreasing NO3- concentrations and CH4 emission in brackish ecosystems with increasing agricultural activities where the native microbial community is incapable of efficient denitrification.

Wetscapes: Restoring and maintaining peatland landscapes for sustainable futures.

Peatlands are among the world's most carbon-dense ecosystems and hotspots of carbon storage. Although peatland drainage causes strong carbon emissions, land subsidence, fires and biodiversity loss, drainage-based agriculture and forestry on peatland is still expanding on a global scale. To maintain and restore their vital carbon sequestration and storage function and to reach the goals of the Paris Agreement, rewetting and restoration of all drained and degraded peatlands is urgently required. However, socio-economic conditions and hydrological constraints hitherto prevent rewetting and restoration on large scale, which calls for rethinking landscape use. We here argue that creating integrated wetscapes (wet peatland landscapes), including nature preserve cores, buffer zones and paludiculture areas (for wet productive land use), will enable sustainable and complementary land-use functions on the landscape level. As such, transforming landscapes into wetscapes presents an inevitable, novel, ecologically and socio-economically sound alternative for drainage-based peatland use.

Anaerobic oxidization of methane in a minerotrophic peatland: enrichment of nitrite-dependent methane-oxidizing bacteria.

The importance of anaerobic oxidation of methane (AOM) as a methane sink in freshwater systems is largely unexplored, particularly in peat ecosystems. Nitrite-dependent anaerobic methane oxidation (n-damo) was recently discovered and reported to be catalyzed by the bacterium "Candidatus Methylomirabilis oxyfera," which is affiliated with the NC10 phylum. So far, several "Ca. Methylomirabilis oxyfera" enrichment cultures have been obtained using a limited number of freshwater sediments or wastewater treatment sludge as the inoculum. In this study, using stable isotope measurements and porewater profiles, we investigated the potential of n-damo in a minerotrophic peatland in the south of the Netherlands that is infiltrated by nitrate-rich ground water. Methane and nitrate profiles suggested that all methane produced was oxidized before reaching the oxic layer, and NC10 bacteria could be active in the transition zone where countergradients of methane and nitrate occur. Quantitative PCR showed high NC10 bacterial cell numbers at this methane-nitrate transition zone. This soil section was used to enrich the prevalent NC10 bacteria in a continuous culture supplied with methane and nitrite at an in situ pH of 6.2. An enrichment of nitrite-reducing methanotrophic NC10 bacteria was successfully obtained. Phylogenetic analysis of retrieved 16S rRNA and pmoA genes showed that the enriched bacteria were very similar to the ones found in situ and constituted a new branch of NC10 bacteria with an identity of less than 96 and 90% to the 16S rRNA and pmoA genes of "Ca. Methylomirabilis oxyfera," respectively. The results of this study expand our knowledge of the diversity and distribution of NC10 bacteria in the environment and highlight their potential contribution to nitrogen and methane cycles.

A microbial consortium couples anaerobic methane oxidation to denitrification.

Modern agriculture has accelerated biological methane and nitrogen cycling on a global scale. Freshwater sediments often receive increased downward fluxes of nitrate from agricultural runoff and upward fluxes of methane generated by anaerobic decomposition. In theory, prokaryotes should be capable of using nitrate to oxidize methane anaerobically, but such organisms have neither been observed in nature nor isolated in the laboratory. Microbial oxidation of methane is thus believed to proceed only with oxygen or sulphate. Here we show that the direct, anaerobic oxidation of methane coupled to denitrification of nitrate is possible. A microbial consortium, enriched from anoxic sediments, oxidized methane to carbon dioxide coupled to denitrification in the complete absence of oxygen. This consortium consisted of two microorganisms, a bacterium representing a phylum without any cultured species and an archaeon distantly related to marine methanotrophic Archaea. The detection of relatives of these prokaryotes in different freshwater ecosystems worldwide indicates that the reaction presented here may make a substantial contribution to biological methane and nitrogen cycles.

New anaerobic, ammonium-oxidizing community enriched from peat soil.

Anaerobic ammonium-oxidizing (anammox) bacteria have been recognized as an important sink for fixed nitrogen and are detected in many natural environments. However, their presence in terrestrial ecosystems has long been overlooked, and their contribution to the nitrogen cycling in natural and agricultural soils is currently unknown. Here we describe the enrichment and characterization of anammox bacteria from a nitrogen-loaded peat soil. After 8 months of incubation with the natural surface water of the sampling site and increasing ammonium and nitrite concentrations, anammox cells constituted 40 to 50% of the enrichment culture. The two dominant anammox phylotypes were affiliated with "Candidatus Jettenia asiatica" and "Candidatus Brocadia fulgida." The enrichment culture converted NH(4)(+) and NO(2)(-) to N(2) with the previously reported stoichiometry (1:1.27) and had a maximum specific anaerobic ammonium oxidation rate of 0.94 mmol NH(4)(+)·g (dry weight)(-1)·h(-1) at pH 7.1 and 32°C. The diagnostic anammox-specific lipids were detected at a concentration of 650 ng·g (dry weight)(-1), and pentyl-[3]-ladderane was the most abundant ladderane lipid.

Zero methane emission bogs: extreme rhizosphere oxygenation by cushion plants in Patagonia.

• Vascular wetland plants may substantially increase methane emissions by producing root exudates and easily degradable litter, and by providing a low-resistance diffusion pathway via their aerenchyma. However, model studies have indicated that vascular plants can reduce methane emission when soil oxygen demand is exceeded by oxygen released from roots. Here, we tested whether these conditions occur in bogs dominated by cushion plants. • Root-methane interactions were studied by comparing methane emissions, stock and oxygen availability in depth profiles below lawns of either cushion plants or Sphagnum mosses in Patagonia. • Cushion plants, Astelia pumila and Donatia fascicularis, formed extensive root systems up to 120 cm in depth. The cold soil (< 10°C) and highly decomposed peat resulted in low microbial activity and oxygen consumption. In cushion plant lawns, high soil oxygen coincided with high root densities, but methane emissions were absent. In Sphagnum lawns, methane emissions were substantial. High methane concentrations were only found in soils without cushion plant roots. • This first methane study in Patagonian bog vegetation reveals lower emissions than expected. We conclude that cushion plants are capable of reducing methane emission on an ecosystem scale by thorough soil and methane oxidation.

Methanotrophic symbionts provide carbon for photosynthesis in peat bogs.

Wetlands are the largest natural source of atmospheric methane, the second most important greenhouse gas. Methane flux to the atmosphere depends strongly on the climate; however, by far the largest part of the methane formed in wetland ecosystems is recycled and does not reach the atmosphere. The biogeochemical controls on the efficient oxidation of methane are still poorly understood. Here we show that submerged Sphagnum mosses, the dominant plants in some of these habitats, consume methane through symbiosis with partly endophytic methanotrophic bacteria, leading to highly effective in situ methane recycling. Molecular probes revealed the presence of the bacteria in the hyaline cells of the plant and on stem leaves. Incubation with (13)C-methane showed rapid in situ oxidation by these bacteria to carbon dioxide, which was subsequently fixed by Sphagnum, as shown by incorporation of (13)C-methane into plant sterols. In this way, methane acts as a significant (10-15%) carbon source for Sphagnum. The symbiosis explains both the efficient recycling of methane and the high organic carbon burial in these wetland ecosystems.

A Novel Laboratory-Scale Mesocosm Setup to Study Methane Emission Mitigation by Sphagnum Mosses and Associated Methanotrophs.

Degraded peatlands are often rewetted to prevent oxidation of the peat, which reduces CO2 emission. However, the created anoxic conditions will boost methane (CH4) production and thus emission. Here, we show that submerged Sphagnum peat mosses in rewetted-submerged peatlands can reduce CH4 emission from peatlands with 93%. We were able to mimic the field situation in the laboratory by using a novel mesocosm set-up. By combining these with 16S rRNA gene amplicon sequencing and qPCR analysis of the pmoA and mmoX genes, we showed that submerged Sphagnum mosses act as a niche for CH4 oxidizing bacteria. The tight association between Sphagnum peat mosses and methane oxidizing bacteria (MOB) significantly reduces CH4 emissions by peatlands and can be studied in more detail in the mesocosm setup developed in this study.